D609 对Neuro-2a 脑神经瘤细胞增殖和细胞周期的影响

目的:探讨小分子化合物D609对脑神经瘤细胞Neuro-2a的生长抑制及诱导细胞周期阻滞的效应,并初步研究其机制。方法:采用CCK-8法检测D609对Neuro-2a细胞的生长抑制作用;利用流式细胞术(FACS)检测D609处理对细胞周期进程的影响;利用免疫印迹实验(Western blot)检测不同浓度的D609处理后,细胞裂解液中细胞周期蛋白抑制因子p27的表达水平。结果:CCK-8的实验结果显示,加入150μmol/L D609处理72小时后,细胞生长受到明显地抑制,且伴有剂量依赖效应;流式细胞术的结果表明,D609处理使细胞周期阻滞在G0/G1期;免疫印迹的结果表明药物处理提升了p27的表达,且随药物浓度升高其表达亦增强。结论:D609可以有效地抑制Neuro-2a细胞的生长;进一步研究表明药物处理可以提升p27的表达水平并可以诱导将细胞阻滞在G0/G1期。因此,此研究将为脑神经瘤的治疗提供借鉴。

The Effect of D609 on Proliferation and Cell Cycle in Neuro-2a Neuroblastoma Cells

Objective:To explore the effect of D609 on cell growth and cell cycle arrest in Neuro-2a cell line, and study the relevant mechanism.Methods:The direct effects of D609 in inhibiting the growth of Neuro-2a cells was examined by CCK-8 assay and the dose-dependent effects was also observed in this assay. Subsequently, we investigated the effects of D609 on cell cycle progression by FACS. Moreover, the expression level of p27, which is a cell-cycle regulatory protein inhibitor, was also detected by western blot when treated with various concentration of D609.Results:CCK-8 assay indicated that D609 (150 umol/L)showed significant concentration-dependent effects in inhibiting Neuro-2a cells with treatment for 72 hours. In FACS assay, D609 induced cell cycle arrest as shown bythe increased percentage of cells in G0/G1 phase. These observations were accompanied by up-regulation of cell-cycle regulatory protein inhibitorp27, which was examined by western blot.Conclusion:D609 could inhibit the growth of Neuro-2a effectively. Moreover, the drug treatment elevated the expression of p27 and further induced G0/G1 arrest. In conclusion, the research will provide useful information for treating neuroblastoma.